Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 57, Issue 25, Pages 7458-7462Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201801016
Keywords
biomolecular NMR; dynamic nuclear polarization; magic-angle spinning; solid-state NMR spectroscopy
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Funding
- CNRS
- EQUIPEX [ANR-10-EQPX-47-01]
- Agence Nationale de la Recherche [ANR15-CE29-002202]
- European Union [317127]
- MC incoming fellowship (REA grant) [661175]
- Fondazione con il Sud [2011-PDR-19]
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Dynamic nuclear polarization (DNP) is a powerful way to overcome the sensitivity limitation of magic-angle-spinning (MAS) NMR experiments. However, the resolution of the DNPNMR spectra of proteins is compromised by severe line broadening associated with the necessity to perform experiments at cryogenic temperatures and in the presence of paramagnetic radicals. High-quality DNP-enhanced NMR spectra of the Acinetobacter phage 205 (AP205) nucleocapsid can be obtained by combining high magnetic field (800MHz) and fast MAS (40kHz). These conditions yield enhanced resolution and long coherence lifetimes allowing the acquisition of resolved 2D correlation spectra and of previously unfeasible scalar-based experiments. This enables the assignment of aromatic resonances of the AP205 coat protein and its packaged RNA, as well as the detection of long-range contacts, which are not observed at room temperature, opening new possibilities for structure determination.
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